Traumatic injury to the central nervous system causes acute neuronal death and surviving injured neurons do not readily regenerate their axons, leading to permanent functional loss. Nanomaterial-based drug delivery systems provide potential for encouraging axon regrowth from specific neurons. In the present study, we analyze the mechanisms of cellular uptake of surface functionalized nanospheres. We used -COOH and –NH2 surface functionalized nanospheres to study the mechanism of cellular uptake in B35 and PC12 cells. The highest dose of –COOH surface functionalized nanospheres showed more perinuclear concentration whereas –NH2 surface functionalized nanospheres showed both nuclear and perinuclear uptake in both B35 and PC12 cells. Also, we found that the –NH2 surface functionalized nanospheres were endocytosed through clathrin as well as caveolin mediated endocytosis in PC12 and B35 cells. In future, we will also investigate the mechanisms of cellular uptake of surface functionalized nanospheres in corticospinal tract neurons. Together, these results will test the feasibility of functionalized nanocarriers for targeted drug delivery to encourage axon regeneration following nervous system damage.